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Welcome & Introduction Occupational Health Occupational Health & Safety Service & Safety Service Samantha Dainty – Biological Safety Adviser Biological and GM safety Agenda Biosafety and Biosecurity • Legislation relating to biosafety • Biosafety and Biosecurity Management Standard • Classifying biological agents • Risk assessment • Notifications and licensing • Containment and controls • Emergency Planning

Genetic Modification • GM Management Standard • GM risk assessment and peer review process

Biological Agents (COSHH) Occupational Health Other legislation Occupational Health & Safety Service & Safety Service

Micro-organisms Human endoparasites zoonoses Toxins

SAPO Schedule 5 Human and animal samples animal pathogens

Environmental samples

Allergens GM Plant health order

Biosafety and Biosecurity Biosafety and Biosecurity Occupational Health Occupational Health Management Standard & Safety Service Management Standard & Safety Service

The standard (duty) • How we will comply with Biosafety legislation

• Who is responsible for How to meet putting the standards the standard Who’s doing into action what

• Applies to everyone at all levels

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Classification of Biological Agents Classification of Biological Agents Occupational Health Occupational Health COSHH & Safety Service SAPO & Safety Service

Human pathogens Or self-classify Animal pathogens Appendix 1 or zoonoses (Schedule 1 of SAPO)

Group Description Examples Group Description Examples HG1 Unlikely to cause human disease B. subtilis, E. coli (K12 and BL21 strains) Disease-producing organisms which are SAPO1 enzootic (native in animals in this country) Can cause human disease and may be a Influenza virus, Listeria Monocytogenes, Adenovirus,, and do not produce notifiable disease. hazard to employees E. coli spp, S. aureus Disease-producing organisms which are Trypanosoma Bruciae, Bovine leucosis virus HG2* Unlikely to spread to community either exotic or produce notifiable disease, Usually effective prophylaxis or SAPO2 but have a low risk of spread from the treatment laboratory. Can cause severe human disease and may be Pandemic influenza virus, HIV, HBV, HCV, E. coli Disease-producing organisms which are Bacillus anthracis, Vesicular stomatitis virus a serious hazard to employees 0157, M. tuberculosis, either exotic or produce notifiable disease HG3* May spread to community SAPO3 and have a moderate risk of spread from Often effective prophylaxis or treatment the laboratory. Causes severe human disease and is a Pandemic influenza virus, Variola virus, Ebola virus Disease-producing organisms which are Avian influenza virus, Foot and mouth disease virus, serious hazard to employees either exotic or produce notifiable disease Rabies virus HG4 Likely to spread to community SAPO4 and have a high risk of spread from the Often no effective prophylaxis or treatment laboratory.

Classification of Biological Agents Classification of Biological Agents Occupational Health Occupational Health Plant Health Order & Safety Service Schedule 5 & Safety Service • Intact Schedule 5 micro-organism Plant Health Schedules 1-6 & 8 • Genetically modified but retain the ability to cause serious harm to (England) Order health • Nucleic acid derived from a Schedule 5 micro-organism that can encode Group Description infectious and/or replication competent forms of the micro-organism • Nucleic acid sequence derived from a Schedule 5 microorganism which Plant pests which shall not be landed in or spread within England. Schedule 1 when inserted into any other living organism alters or enhances that Relevant material which may not be landed in or moved within England Schedule 2 organisms's ability to cause serous harm to human or animal health if that material is carrying or infected with plant pests • any quantity of botulinum or Clostridium perfringens Relevant material which may not be landed in England if that material enterotoxin/epsilon toxin Schedule 3 originates in certain third countries • more than 5 mg of any other schedule 5 toxin Restrictions on the landing in and movement within England of relevant Schedule 4 material • Any nucleic acid sequence coding for a Schedule 5 toxin or a genetically Relevant material from a third country for which a phytosanitary modified micro-organism containing such a sequence Schedule 5 certificate may be required Prohibitions on the landing in and movement within England of relevant Schedule 6 material without a plant passport Swiss plant passports Schedule 8

Notifications and Licensing Occupational Health BioCOSHH Risk Assessment Occupational Health & Safety Service & Safety Service Pathogen and toxin registration

Requirements for work with biological agents according to their classification

Specified Animal ACDP Hazard Group Plant Health Order Materials that may Schedule 5 Classification Pathogen Controlled or prohibited plants, contain biological Review every Advice: plant products or plant pests Anti terrorism, Crime & Security • Biological Safety 1 2 3 4 1 2 3 4 under Schedules 1-6 agents Act 2 years (at Supervisors Assessment Risk Assessment least) • School Safety CL CL CL CL CL CL CL Compliance with the See OHSS H&S Guidance Biosecurity policy in Officers Containment 1 2 3 4 2 3 4 conditions of the licence 301.3 consultation with CTSA • Biological Safety Pathogen Pathogen Pathogen registration if known/very Registration Pathogen registration likely to contain ACDP HG2-4, SAPO Pathogen registration Advisor registration registration 2-4 etc.

*Onl HSE SAPO/IAPO Notifications or licences depending y 1st Notification Plant Health licence on nature of the sample CTSA notification use notification licences HG3, SAPO3, PHO licences, Schedule 5 = Peer review by Biosafety Committee

AND Let us know when your work has ceased

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Routes of exposure Occupational Health Infectious Dose Occupational Health & Safety Service & Safety Service

Mucous Membranes: Exposure of mucous membranes of the Amount of pathogenic organisms that will cause infection in susceptible subjects eyes, nose and mouth through splashes, splatter or aerosols. Ingestion: Swallowing through mouth Agent Dose Route pipetting, eating, drinking or Inhalation: smoking in the lab. Tuberculosis 10 Inhalation Breathing in respirable sized aerosols (<5μm), centrifuge leaks, Influenza A2 790 Inhalation spills, pipetting, etc. S.typhi 105 Ingestion Percutaneous/injection: Vibrio cholerae Cholera Through intact or non-intact skin via 108 Ingestion NEEDLESTICK, puncture with contaminated sharp 8 object, animal scratch or bite, through wounds E.coli 10 Ingestion abrasions, or eczema. E.coli 0157 10-100 Ingestion

Contact (indirect transmission): Shigella 10 Ingestion Via mucous membranes or non-intact skin from hands that have been in contact with a contaminated surface Polio virus 1 2 Ingestion (i.e. benches, phones, computers, equipment handles) or by failure to wash hands after working

Quantity and Concentration Occupational Health Who is at risk? Occupational Health & Safety Service & Safety Service

Concentration, centrifugation, filtration

Small scale

Higher risk? Anyone at increased risk? Supervision/access Highly concentrated substances can be: - Immunocompromised control - Asthma Shared spaces – • More infectious - Dermatitis Severity of exposure • who else needs to be Higher aerosol potential informed? • Emergencies require specialist spill clean up Experience – impacts likelihood of Medium scale Large scale • Waste inactivation issues exposure • Higher risk

Risk Estimation Occupational Health Controls – Containment labs Occupational Health & Safety Service & Safety Service

Containment level 1 - hazard group 1 Containment level 2 - hazard group 2 Frequency Route of Infectious dose Containment level 3 - hazard group 3 exposure

Containment level 2 for when there is no intention to deliberately propagate or Nature of concentrate a biological agent but there are organism/ uncertainties about the presence of HG2, HG3 hazard group or HG4 in the sample.

concentration Containment level 3 or 4 where appropriate if it known or suspected that such a level is necessary even if there is no intention to deliberately propagate a HG3 or HG4 biological agent

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Controls – Containment labs Occupational Health Containment level 1 Occupational Health & Safety Service & Safety Service

Facilities • Hand wash sinks Equipment • Bench surfaces (and floors) impervious to water and System of work chemicals. Easy to clean Waste and disinfection • Autoclave on site (GM) • Suitable protective clothing Animal containment • Specified disinfection procedures • Inactivation of GMMs by a validated means • Written records of staff training

Containment level 2 Occupational Health Containment level 3 Occupational Health & Safety Service & Safety Service

• Isolation of laboratory suite • Procedures involving infectious • Lab sealable for fumigation aerosols must be contained within an • Entry via airlock MSC (or similar) • Negative pressure • Access restriction • HEPA filtered extract • Biohazard sign on door • Procedures with infectious materials • Autoclave in the building must be contained within MSC or • Inactivation of all biological materials similar by a validated means • Access restriction • Safe storage of biological materials • Biohazard sign on door • Autoclave in the lab • Inactivation of all biological materials by a validated means • Safe storage of biological materials

SAPO Containment labs Occupational Health Containment labs Occupational Health & Safety Service & Safety Service

Plant Health Licences – You must meet the conditions of the licence Containment level 2 – SAPO group 2 Containment level 3 – SAPO group 3

Schedule 5 – Security control measures as advised by CTSA and a Biosecurity Policy

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Multiuser labs Occupational Health Controls – COSHH Hierarchy Occupational Health & Safety Service & Safety Service

Hierarchy of control selection • All users must comply with the requirements of the lab containment level regardless of the hazard group of the organism they are working on Attenuated strains • Good communication required Auxotrophs ‘lab’ strains

Microbiological Safety Cabinets Occupational Health Microbiological Safety Cabinets Occupational Health & Safety Service & Safety Service

Correct Operation is essential for containment

• Do not obstruct grill • Bunsen burners, centrifuges and walk- past all disrupt air flow • MSCs do not sufficiently vent harmful chemicals. Avoid volatile chemicals – methanol, acetic acid, mercaptoethanol • Caustic material that vaporize can attack aluminum separators in the HEPA filter • Statutory requirement – annual LEV testing

Containing aerosols in centrifuges Occupational Health Animal Containment Occupational Health & Safety Service & Safety Service • Use biosafety rotors for biological agents (CL2+) • Open rotor, buckets, tubes inside MSC • Do not open centrifuge immediately after a breakage/spillage – let aerosols settle first • Disinfect after spillages

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Administrative Controls Occupational Health Personal Protective Equipment Occupational Health & Safety Service & Safety Service

Gloves – select the right type training Access control

Lab coats

Eye protection

Signs Systems of work Respiratory protection – face fit testing

Immunisation Occupational Health Health Surveillance Occupational Health & Safety Service & Safety Service • We are required by law to keep a list of those working with Area Activity ** Required Recommend

Clinical Patients/clinical materials contact – exposure prone procedures Hep B* hazard group 3 biological agents

Patients/clinical materials contact – non exposure prone procedures Hep B Laboratory Working with unscreened human blood or tissue Hep B • Asthmagens Working with screened human blood or tissue Hep B • Allergens Working with novel human cell lines from uncontrolled sources Hep B • Certain pathogens e.g. TB Working with established human cell lines from controlled sources Hep B NB Has the cell line been screened? Pre 2010 ATCC? Working with non human material Health surveillance is required if all the following criteria are met:

Agricultural / Handling soil or plant material Tetanus Horticultural • There is an identifiable disease/adverse health effect and evidence of a link Animal Working with colony bred animals Tetanus with workplace exposure Technicians • It is possible the disease/health effect may occur Working with wild caught primates if not conditioned Rabies, Hep A Tetanus • There are valid techniques for detecting early signs of the disease/health effect • These techniques do not pose a risk to employees

Principals of Good Microbiological Practice Transport of Biological Samples Occupational Health Occupational Health & Safety Service & Safety Service

Aseptic technique • Between labs - Secondary containment • Between university sites:

housekeeping

Aerosol minimising techniques

• Between Organisations – Sign up for the training course!

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Waste Inactivation - autoclaving Occupational Health Waste Inactivation - autoclaving Occupational Health & Safety Service & Safety Service • Waste must be inactivated by a validated means – autoclave in preference to disinfection Contaminated with • CL2 – before removal from the site biological agents or GM. No chemicals • CL3 – before removal from the facility (residues OK)

• Applies to ALL GM samples Energy from monitoring waste/deep landfill Contaminated with biological agents or High temperature GM. Can’t be incineration autoclaved Take the tops off your tubes!!!

Not contaminated with biological agents but looks like lab waste (e.g. • But…..do not autoclave samples containing radioactive or gloves from a chemistry hazardous chemicals lab) No chemicals

Sharps Occupational Health Waste Inactivation - disinfectants Occupational Health & Safety Service & Safety Service

A specified disinfection procedure is required for all work with biological agents and GM microorganisms

Practice Disinfectant Bacteria Bacterial Fungi Enveloped Non-enveloped Prions Comments

spores viruses viruses Bad Use gloves and Never Don’t dispose of Don’t dispose of Don’t transfer Phenolic YES NO YES YES Limited NO Toxic never resheath resheath sharps in clinical sharps in ordinary used sharps to needles needles waste bags waste bins other workers Hypochlorites YES YES Limited YES YES YES Toxic/Corrosive

70% Alcohol YES NO NO YES Limited NO Flammable Irritant/allergen,

Aldehydes* YES YES YES YES YES NO glutaraldhyde resistance Practice Practice bacteria CL3 level - very toxic,

*Formaldehyde YES YES YES YES YES NO Good Good Good Good need to seal the lab! Dispose of sharps Take disposal bin Dispose of bins Treat all Dust irritant, limited Peroxygen solution life, corrosive to immediately after to sharps on reaching max contaminated sharps YES YES YES YES YES NO use in a sharps bin level (marked) as hazardous (Virkon) metals

Waste Inactivation - disinfectants Occupational Health Waste Inactivation - disinfectants Occupational Health & Safety Service & Safety Service

Validation! • some disinfectants can be ‘inactivated’ How do you know if disinfection has been under different conditions successful? • presence of ‘organic material’ - • data from manufacturer – kills 100% of all compensate known germs! • avoid disinfectant combinations • data from other users / labs • limit age of disinfectant solution (fresh) • • ‘in-use’ testing use at correct dilution (‘in-use’ concentration) • exposure time • COSHH properties; compatibility with hardware

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Emergency Planning Occupational Health Emergency Planning Occupational Health & Safety Service & Safety Service Spills Needlestick/Contaminated Sharps absorbent towels SOP absorbant granules lab coat • disinfectant Action after exposure to blood borne viruses – Occupational Health • Risk assessment • Procedures for potential exposure to HepB (if unvaccinated), sealable bin overshoes HepC and HIV (autoclavable) • counselling

goggles autoclave bags Mask biohazard (HEPA gloves tape gloves filtered) (heavy duty)

Definition Occupational Health Occupational Health & Safety Service & Safety Service Any technique which alters the genetic material of an organism using a method that would not occur by natural mating or Genetic Modification recombination

Gene deletions or insertion of multiple copies of a gene if brought about by artificial means

e.g. Insertion of nucleic acid, produced outside an organism into any virus, plasmid or other vector then incorporation into a host organism in which it does not occur naturally and a capable of continued propagation.

e.g.. Direct introduction into an organism of heritable genetic material prepared outside the organism, including micro-injection and micro-encapsulation.

Exemptions Occupational Health GM risks: The Unknown Occupational Health & Safety Service & Safety Service

Mutagenesis (eg x-rays, chemicals)

Synthetic nucleotides

Self cloning organisms

“Natural” transformation

Hybridoma’s

Humans and human embryos - IVF

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Genetic Modification Management GM risks: Public Perception Occupational Health Occupational Health & Safety Service Standard & Safety Service

• How we will comply with the Contained Use regulations

• Who is responsible for putting the standards into action

• Applies to everyone at all levels

We have to protect ourselves/work from this controversy by demonstrating the highest level of containment of GMO – “zero risk” of escape……

Risk Assessment Occupational Health Risk assessment for……. Occupational Health & Safety Service & Safety Service

Host GM cells

Genetic material Gene cDNA

Vector

Use this to help! Occupational Health Occupational Health & Safety Service Waste Inactivation & Safety Service

Health and safety inspectors seek to secure compliance with the law and All GM waste must be inactivated by a validated may refer to this guidance as means illustrating good practice Autoclaves in preference to disinfectants

Must be described in the GM risk assessment

Any deviation from this must be in discussion with the Biological Safety Adviser - Consultation with HSE

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Controls and Activity Class Occupational Health Control Selection and Activity Class Occupational Health & Safety Service & Safety Service

Containment necessary to control Risk Classification the risk HG1 - GM Class 1 – CL1 Level 1 Class 1 HG2 - GM Class 2 – CL2 HG3 - GM Class 3 – CL3 Level 1 with the addition of measures from level 2 Or Class 2 Level 2 (without additional measures) BUT an HG1 organism could become a GM class 2 depending on how it’s been modified Level 2 with the addition of measures from Level 3 Class 3 Or Hazard group rating sets the “base level” then depending on the Level 3 (without additional measures) modification the organism may be elevated to a higher risk group based

Level 3 with the addition of measures on the modification from Level 4 Or Class 4 Level 4 (with or without additional measures)

Control Selection and Activity Class Occupational Health Peer Review Procedure Occupational Health & Safety Service & Safety Service

Class Criteria for Classification 1  3rd generation (or safer) vector system All criteria must be  Self inactivating (SIN) LTR met to justify the  Non harmful insert assignment of  Exclusion of sharps Class 1  Low viral titres  1st/2nd generation vector system 2  Vectors containing X-protein expressing forms of the woodchuck If at least one of hepatitis B virus post-transcriptional regulatory element the criteria applies  Harmful insert the activity will be  Use of sharps required* Class 2  High viral titres and/or aerosol generating procedures warranting the £981! use of a microbiological safety cabinet**

*’Whilst ‘control of sharps’ is not one of the specified control measures in the containment tables, other containment level 2 measures are deemed necessary to facilitate their control (e.g. restricted access to authorised and trained personnel; written training records; and the use of gloves) therefore necessitating a classification of Class 2.’ (SACGM compendium of guidance). **Does not apply if the use of the cabinet is solely for purpose of ensuring the sterility of the work.

Peer Review Procedure Occupational Health And finally……… Occupational Health & Safety Service & Safety Service

Changes to: • Host Let us know when your GM work has ceased • Vector • Insert • Personnel AND • Location • Closing date The samples have been destroyed or transferred elsewhere

£734! We must tell HSE when activity class 2/3 projects have ceased We can submit a transfer notification to HSE if you move to another university We need to keep the GM register up to date

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Any Questions?

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